Culture of bone marrow mesenchymal stem cells plus allogenic decalcified bone matrix in the knee cavity of rabbits for tissue engineered cartilage

doi:10.3969/j.issn.2095-4344.2013.24.001

Abstract

Abstract:

BACKGROUND: It remains controversial how to repair articular cartilage defects and achieve a good long-term efficacy using tissue engineering methods.
OBJECTIVE: To investigate the feasibility of culturing rabbit bone marrow mesenchymal stem cells in combination with allogenic decalcified bone matrix in the knee cavity of rabbits for constructing tissue engineered cartilage.
METHODS: Bone marrow mesenchymal stem cells were isolated using adherence screening method and cultured in complete Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12. The third generation was induced to chondrocytes. Allogenic decalcified bone matrix, made from rabbit’s ilium and vertebral bone, was combined with the induced bone marrow mesenchymal stem cells in vitro for 1 day. The cell-scaffold composites wrapped by fascia were cultured in the left knee cavities of adult rabbits (cell-scaffold group, n=15) while simple homologous decalcified bone matrix wrapped by fascia were cultured in the right knee cavities (scaffold group, n=15). The specimens were harvested at 4, 8, and 12 weeks after culture for gross and histological observations. Histological observation was conducted using hematoxylin-eosin staining, toluidine blue staining and immunohistochemical staining.
RESULTS AND CONCLUSION: At 4 and 8 weeks after culture, the mean absorbance value was respectively 0.263±0.031 and 0.340±0.052 for the cell-scaffold group and 0.147±0.027 and 0.165±0.030 for the scaffold group, with significant differences between two groups (P < 0.05). At 12 weeks, the mean absorbance value of specimens in the cell-scaffold group was 0.362±0.037, and the samples exhibited similar appearance of normal cartilage and positive immunohistochemical reaction of type II collagen. However, the scaffold degraded in the scaffold group. At 12 weeks, hematoxylin-eosin staining revealed increased chondrocytes and mostly absorbed decalcified bone matrix scaffold in the cell-scaffold group while toluidine blue staining showed metachromasia matrix dyed red or purple. Results indicate that tissue engineered cartilage can be induced by culturing rabbit bone marrow-derived mesenchymal stem cells combined with allograft decalcified bone matrix in the knee cavity of rabbit.

Key words: tissue construction, cartilage tissue construction, bone marrow mesenchymal stem cells, cytoskeletal proteins, allograft, decalcified bone matrix, tissue engineering, chondrocytes, rabbits, provincial grats-supported paper

CLC Number:

R318

Figures/Tables 7

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